The proposed research program is concerned with the structure, metabolism and function of protein-acid mucopolysaccharide complexes in normal individuals and patients with connective tissue disease. Emphasis will be placed on the anabolism and catabolism of the protein- chondroitin sulfate-keratan sulfate-complex. Studies concerned with the biosynthesis of the carbohydrate portion of the complex will focus on the possible involvement of the nucleotide and/or isoprenoid phospho di- and oligosaccharides as immediate precursors of growing polysaccharide chains. Catabolic studies will emphasize the purification of mammalian glycosidases and sulfatases required for the degradation of keratan sulfate. These studies will involve attempts to purify glycosidases by affinity column chromatography. If these studies are successful, additional studies will be initiated to elucidate the nature of the linkage region(s) between keratan sulfate and polypeptide core. The action of one of these enzymes (Beta-galactosidase) will be examined for its ability to "correct" the abnormal accumulation of keratan sulfate (and G sub Ml ganglioside) in cultures of G sub Ml gangliosidosis and Morquio's syndrome fibroblasts. In separate but related studies, the substituents of the protein- chondroitin sulfate-keratan sulfate complex found in normal articular cartilage and those from patients with connective tissue disease will be studied to determine 1) if antigenic alterations of the cartilage matrix or its degradation products occur, 2) and if an autoimmune response may result from prolonged exposures of tissue to "sensitized" cartilage components.